The Annals of the University Dunarea de Jos of Galati

Fascicle VI – Food Technology (2018), 42(2), 49-58

ORIGINAL RESEARCH PAPER

NANOCOMPOSITE EDIBLE COATING FROM CASSAVA STARCH,

STEARIC ACID AND ZnO NANOPARTICLES TO MAINTAIN QUALITY
OF FRESH-CUT MANGO CV. ARUMANIS

SRI YULIANI1, ATA ADITYA WARDANA2*, BAYU MEINDRAWAN2, NUGRAHA EDHI

SUYATMA3, TIEN R. MUCHTADI3

1
Indonesian Center for Agricultural Postharvest Research and Development, Bogor,
16114, Indonesia
2
Department of Food Technology, Faculty of Engineering, Bina Nusantara
University, Jakarta, 15143, Indonesia
3
Department of Food Science and Technology, Bogor Agricultural University, Bogor,
16680, Indonesia
*Corresponding author: ataaditya@gmail.com, ata.wardana@binus.edu

Received on 22nd March 2018

Revised on 4th July 2018

Fresh-cut mango is one of perishable food products due to its high respiration,
transpiration and microbial decay rate. In this study, generation of a new edible coating
was chosen as an alternative method for preserving the quality attributes of fresh-cut
mango. The objective of this study was to evaluate the effects of nanocomposite edible
coating from cassava starch (SC), stearic acid (SA) and/or ZnO nanoparticles (NP-ZnO)
on the quality of fresh-cut mango cv. Arumanis, stored for 12 days at 8°C. The
formulations of 20 g/L SC, 6 g/L SA, and variable NP-ZnO (0, 1, 2% w/w of SC) were
utilized to coat the fresh-cut mango. The result revealed that the treatment of
nanocomposite edible coating was able to maintain the quality of fresh-cut mango
during the storage periods. The SC, SA, and 2% NP-ZnO formulation was the most
effective in reducing weight loss and microbial counts. Our study suggests that the
application of nanocomposite edible coating provides a desirable method to maintain
storage quality and to improve fresh-cut mango postharvest life.

Keywords: mango, fresh-cut, nanocomposite, ZnO nanoparticles, edible coating

Introduction
Mango (Mangifera indica L.) is one of the tropical and climacteric fruits, a rich
source of minerals, carotenoids, vitamins A, and B, with a wide demand on the
international fruit market (Chiumarelli et al., 2010). The top five mango-producing
countries from 2003 to 2005 were India (38.6%), followed by China (12.9%),
Thailand (6.2%), Mexico (5.5%) and Indonesia (5.3%) (FAOSTAT, 2007). In
Indonesia, mango cv. Arumanis is the most popular mango variety to consume with
a weight of about 300–500 g per fruit. The increasing consumer demand for healthy,
fresh, and ready-to-eat fruit is a current driving force for the expansion of the fresh-
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cut mango market. However, fresh-cut mango has a short period of shelf life due to
its lack of preparation, which stimulates respiration and transpiration, membrane
deterioration, and supports microbial growth (Laurila and Ahvenainen 2002; Rojas
et al., 2009). Hence, alternative methods are highly needed to maintain the quality
and to prolong the fresh-cut mango storage periods.
Edible coating is one of the recommended methods to inhibit deterioration of fresh-
cut products. It has a role as semipermeable barrier, which makes it able to inhibit
moisture and solute migration, gas exchange, respiration and oxidative reaction rates,
as well as suppress physiological disorders of fresh-cut fruits (Robles-Sánchez et al.,
2013). Moreover, edible coatings are also utilized as carrier of food additives
including antimicrobial agents (Shi and Gunasekaran, 2008; Suyatma et al., 2014).
Among biopolymers matrices, starch is still one of the most promising polymers due
to its availability and low cost (Ma et al., 2009). A hydrophilic single coating has a
good barrier property to gases but is weak on water vapor resistance. To improve the
barrier to water vapor, the incorporation of hydrophobic materials such as long-chain
fatty acids were applied as an alternative (Schmidt et al., 2013).
Nowadays, the utilization of nanoparticles (NP) has gained great attention in
developing nanocomposites. A nanocomposite is a breakthrough of a
microcomposite, which consists of natural polymers as matrix with nanoparticles as
filler. The addition of nanoparticles into biopolymers are able to improve their
characteristics and functional properties including barrier ability, morphology, and
mechanical strength (Sorrentino et al., 2007; Shi and Gunasekaran, 2008; Slavutsky
and Bertuzzi, 2014; Shankar et al., 2015). NP-ZnO is one of the nanoparticle
substances that has received great attention as nano fillers in the food field.
Furthermore, NP-ZnO has been approved as GRAS (Generally Recognized as Safe)
substance by the FDA (Sharon et al., 2010). It also was applied as a source for Zn
supplementation and fortification (Shi and Gunasekaran, 2008). It also was revealed
that NP-ZnO acted as antimicrobial substances. Yu et al. (2009) incorporated NP-
ZnO into the polymers to reduce the growth of microorganisms on the surface of
foodstuffs. Previous studies also found new possibilities of NP-ZnO against Botrytis
cinerea and Penicillium expansum (He et al., 2011), Escherichia coli (Zhang et al.,
2007), and Lactobacillus plantarum (Shi et al., 2012). Thanks to its safety and its
beneficial properties, NP-ZnO was selected in this study to develop a nanocomposite
edible coating. The aim of this work was to evaluate the effects of a nanocomposite
edible coating from cassava starch (SC), stearic acid (SA) and NP-ZnO on the quality
of fresh-cut mango cv. Arumanis, stored for 12 days at 8°C.

Materials and methods

Materials
The materials used in this study was mango (Mangifera indica L.) cv. Arumanis
obtained from mango farmers at Astanajapura, Cirebon (Indonesia). The mangoes
were harvested approximately 90 days after flowering. Cassava starch was purchased
from local starch industry in Ciluar, Bogor (Indonesia). Stearic acid was purchased
from local market and NP-ZnO with an average particle size of 20 ± 5 nm was
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purchased from Wako Pure Chemical Industries Ltd (Japan). In order to obtain good
dispersion Tween 80 (Merck, Germany) was used as emulsifier.
Preparation of nanocomposite coating solution and samples
NP-ZnO (0, 1 and 2% w/w of SC) was dispersed in 500 ml distilled water by using an
ultraturax (1500 rpm, 10 min) to make the nanocomposite edible coating solutions.
An amount of 10 g cassava starch was slowly added and blended under constant
stirring until it was dissolved completely. Subsequently, 75% Tween 80 (w/w of SC;
or 7.5 g) and 30% SA (w/w of SC; or 3 g) were added into the prepared solutions. The
mixture was heated, and stirring was continued until 90oC for 10 min, and then the
mixture was left until it reached room temperature (28-30oC) naturally. Thus, three
different nanocomposite coating formulations were produced, namely NP-ZnO 0%
(SC+ 30% stearic acid + 0% NP-ZnO), NP-ZnO 1% (SC + 30% stearic acid + 1%
NP-ZnO), and NP-ZnO 2% (SC + 30% stearic acid + 2% NP-ZnO).
The fresh-cut mango used as samples were prepared aseptically to minimize
contamination. Mango fruits were selected for their uniformity in weight (350-400
g) and absence of defective spots, cleaned and washed in 200 ppm chlorine solution.
The fruits were peeled manually with a stainless steel knife and sliced from both
sides of the seed. Subsequently, flesh of mangoes were cut in pieces (1.5-2 cm3) and
immersed in 1% CaCl2 water for 5 min. Then the mango pieces were dipped for 1
min in the different nanocomposite solutions and drained. After being air-dried, the
sliced fruits were placed into polyethylene terephthalate plastic trays (11 x 8 cm).
All samples were stored at 8°C for 12 days.
Weight Loss
Weight loss of samples was determined using an analytical balance at the beginning
of the experiment after coating and air-drying, and thereafter at 3, 6, 9 and 12 days
during the storage period. Measurements were performed with three replications.
Weight loss was calculated using equation 1.
Weight loss (%) = (intial mass - final mass)/initial mass x 100% (1)
Firmness
Firmness of the samples was measured by using a texture analyzer instrument (CT
V1.2 Brookfield, USA) with a probe type TA 39 and 4500 g compression load.
Compression was forced in the central zone for each mango cube (1.5-2 cm3). The
firmness of samples was measured as the maximum penetration force (N) recorded
during tissue breakage.
Browning Index
The color of mango samples were measured by using a Chromameter Minolta CR-
300 based on the CIE color system, where color coordinates range from L*= 0
(black) to L*= 100 (white), -a* (greenness) to +a* (redness), and -b* (blueness) to
+b* (yellowness). The values of measurements were converted to a Browning Index
using equation 2 (Ergüneş and Tarhan, 2006).
Browning Index = [100 (x - 0.31)] / 0.17 (2)
where: x = (a* + 1.75L*) / (5.645L* + a* - 0.3012b*)
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Total Microbial Count

Microbial count of samples was determined using the pour plate method and plate
count agar obtained from Merck (Germany) as the medium. A portion of 25 g of
homogenized fresh-cut mangoes were transferred in 225 mL 0.1% peptone water.
Further decimal dilutions were made from this 10-1 dilution. Subsequently, each
sample was inoculated on to the media and incubated at 37°C for 48 h, following the
FDA Bacteriological Analytical Manual (1998). Colony counts were converted into
log cfu/g (colony forming units per g of sample).
Statistical analysis
The quality parameters of fresh-cut mango were analyzed statistically using
completely randomized design. The results were evaluated by analysis of variance
(ANOVA) using SPSS (Statistical Product and Service Solutions) version 16.0 and
compared by Duncan’s Multiple Range Test (DMRT) when needed, used to
represent results at a significance level of p < 0.05.

Results and discussion

Weight Loss
Figure 1 shows that the weight loss of fresh-cut mango tended to increase during
storage. Lin and Zhao (2007) stated that edible coating application on vegetables and
fruits could inhibit gas exchange which ultimately inhibits weight loss. On day 12,
fresh-cut mango (control, without coating) has bigger percentage of weight loss
(5.7%) than mango with coating treatment (1.5-2.6%). Incorporation of 2% NP-ZnO
into nanocomposite (ZnO formulation) resulted in the smallest weight loss.
However, there was no statistical difference between the different NP-ZnO
concentrations (p > 0.05). NP-ZnO is known as a physical barrier for water vapour
and gases (Torabi and Nafchi, 2013). The respiration process affects weight loss due
to its contribution on water loss through decomposition of complex compounds into
CO2 and H2O. A similar result was reported by Marpaung et al. (2015) with
application of 1% starch or 1% pectin and 1% NP-ZnO nanocomposite coating on
fresh-cut salak (Salacca zalacca) fruit, which showed a weight loss about 3% at the
end of storage (day 12). Also here, nanoparticle embedded-coating could surpress
the weight loss change compared to the control sample. This phenomenon is caused
by inhibition of water evaporation rate, respiration and complex compounds
decomposition.
Firmness
The firmness of fresh-cut mango tended to decreased during storage (Figure 2) with
0.16 N to 0.27 N. This result is similar to that observed by Sothornvit and Rodsamran
(2008), where the firmness reduction of fresh-cut mango with combination of film
wrap and modified atmosphere packaging during 6 days storage was 0.10 – 0.35 N.
Figure 2 shows that the different nanocomposite coating treatments did not give any
statistical differences (p < 0.05). It could however be suggested that coating
treatment also does not lead to a negative effect on the firmness of mango.
Chatanawarangoon (2000) stated that decreasing firmness of fresh-cut mango is
 Yuliani et al. / AUDJG – Food Technology (2018), 42(2), 49-58 53

caused by water loss. Decreasing the water content caused decreasing turgor
pressure, rigidity level and firmness of fruit. In addition, during the fruit ripening
process, insoluble pectin (protopectin) changes into soluble pectin. Microbial
activity also causes mango to become soft and watery. Protopectinases secreted by
bacteria could speed up protopectin decomposition into pectin, thereby causing the
fruit tissue to become soft also.

Figure 1. Weight loss of fresh-cut mango during storage. Different letters indicate
statistically significant differences (p < 0.05).

Figure 2. Firmness of fresh-cut mango during storage.

Color Change
The color change of fresh-cut mango was determined using CIE system by
calculating L*(brightness), a* (redness) and b* (yellowness) values. Subsequently,
they were converted into a browning index. The browning reaction happened to all
54 Yuliani et al. / AUDJG – Food Technology (2018), 42(2), 49-58

fresh-cut mango treatments during storage (Figure 3). Enzymatic browning is known
to be caused by polyphenol oxidation. This reaction is catalyzed by the polyphenol
oxidase enzyme, which form a brown melanine compound. Peeling and cutting could
make the contact area between fruit and oxygen become larger, so the activity of
phenolase enzyme is increased. Nanocomposite coating could inhibit the
enhancement of browning index during storage (Figure 3). Solivia-Fortuny and
Martin-Belloso (2003) stated that coating application based on polysaccharides was
able to prevent dehydration, lipid oxidation, browning and decrease respiration rate
by controlling internal CO2 and O2 composition. In this research, nanocomposite
coating significantly surpressed the browning index of fresh-cut mango. Application
of NP-ZnO formulation resulted on the largest reduction of browning index (p <
0.05). Li et al. (2011) found similar result on fresh-cut “Fuji” apple when stored with
NP-ZnO containing packaging. Li et al. (2011) also suggested that the polyphenol
oxidase activity was decreased significantly resulting in lower Browning Index than
control. Moreover, the change of browning index can also be influenced by microbial
activity. Microbes, predominantly molds such as Aspergillus niger, contribute to the
browning reaction. This is indicated by large-sized spots that make the fruit color
become brown or dark.

Figure 3. Browning index of fresh-cut mango during storage. Different letters indicate
statistically significant differences (p < 0.05).

Microbiological analysis
Total microbes in all treatments increased during storage as shown in Figure 4 by
which the initial and the end microbial counts of fresh-cut mango reached 3.03 and
8.38 log cfu/g respectively. Indonesia National Agency of Drug and Food Control
(BPOM) (2009) regulates that the maximum of microbial contamination on fruit is
5.00 log cfu/g. On day 6, nanocomposite coating treatments significantly suppressed
microbial growth below the safe threshold for consumption compared to control
(5.38 ± 0.04 log cfu/g) and NP-ZnO 0% coated mango (5.36 ± 0.05 log cfu/g). This
is due to the nanocomposite NP-ZnO filler which has antimicrobial ability. Several
 Yuliani et al. / AUDJG – Food Technology (2018), 42(2), 49-58 55

mechanisms of NP-ZnO as antimicrobial have been reported, such as i) intracellular

NP-ZnO accumulation, ii) destruction of bacterial cell membranes, iii) H2O2
generation and iv) by releasing Zn2+ ions. E.g., NP-ZnO penetrate into the cells of
bacteria through a hole or protrusion causing membrane disruptions, so that the cell
experienced lysis (Yousef and Danial 2012). Others suggested that the antimicrobial
mechanism could be seen from the interaction of NP-ZnO with phosphorus groups
in DNA causing replicating inactivation and inhibition of enzyme function of
bacteria (Arabi et al., 2012).

Figure 4. Microbial counts of fresh-cut mango during storage. Different letters indicate
statistically significant differences (p < 0.05).

To provide better illustration of the antimicrobial effect of the nanocomposite NP-

ZnO, Figure 5 demonstrates a comparison of all fresh-cut mangoes at the initial day
(day 0) and the end storage (day 12). At the end of storage some microbial colonies
were clearly visible on the surface of the control and NP-ZnO 0% marked by several
dark spots. These are reduced or absent on the NP-ZnO 1% and 2% formulations.

Figure 5. Comparison of fresh-cut mango during storage (day 0 and day 12).
56 Yuliani et al. / AUDJG – Food Technology (2018), 42(2), 49-58

Conclusions
We have developed novel nanocomposite coatings containing cassava starch, stearic
acid and various amounts of NP-ZnO. Application of these nanocomposite coatings
on fresh-cut mango was capable of suppressing the weight loss without negatively
affecting the firmness of the fresh-cut mango during storage at 8oC, for 12 days.
Furthermore, at day 6 of storage nanocomposite coating treatments of NP-ZnO 1%
and 2% suppressed microbial activity below the safe threshold for consumption (5.00
log cfu/g) to 4.95 ± 0.03 and 4.80 ± 0.07 log cfu/g, respectively. At the same time
the mango control (5.38 ± 0.04 log cfu/g) and ZnO 0% coated mango (5.36 ± 0.05
log cfu/g) were not feasible to be consumed with respect to safety. The edible coating
with 2% ZnO (of SC) performed superior in all aspects of the analyses. By
considering our results, the novel nanocomposite coatings represent a possible
alternative to maintain quality of fresh-cut mango with the objective to preserve the
weight loss during storage and to delay the microbial growth.

Acknowledgments
This research was financially supported by the Indonesian Agency for Agricultural
Research and Development, Ministry of Agriculture, through the Consortium Agro-
Nanotechnology, Indonesia 2014 - 2015. In addition, we thank to Prof. Koen
Venema from Maastricht University and Dr. Dave Mangindaan from Bina Nusantara
University for the writing assistance and proof reading the article

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